Mouse NFkB Secreted Alkaline Phosphatase Reporter SEAP - (SEAPorter™) Stable Reporter Cell Line

Product Discontinued

Mouse NFkB Secreted Alkaline Phosphatase Reporter SEAP - (SEAPorter™) Stable Reporter Cell Line Summary

• Description: The RAW reporter cell line can be used for screening of TLR agonists or antagonists as well as inhibitory TLR antibody assay. RAW 264.7 cells are known to respond to most Toll-like receptor (TLR) ligands, which trigger the NF-kB induction and lead to inflammatory cytokine production. RT-PCR tests also shows that RAW 264.7 cells produce all of the TLR mRNAs except for TLR5 (Figure 1). Using a 96-well plate format assay, the RAW cell line has been validated by TLR ligand stimulation in which all of the TLR ligands except for TLR5 ligand activated the cell line (Figure 2).
  
  Contents: 3~4 x 10^6 cells
  Biosafety Level: 2
• Immunogen: The RAW reporter stable cell line is a stably transfected RAW 264.7 cell line which expresses the secreted alkaline phosphatase (SEAP) reporter gene under the transcriptional control of an NF-kB response element.
• Target Species: Mouse
• Specificity: RAW NF-kB/SEAP Reporter Cell Line
• Selection Agent: RAW cell line.
• Growth Properties: Adherent Morphology: Macrophage
• RCL Type: SEAP - (SEAPorter™)
• Reporter Gene: Secreted alkaline phosphatase (SEAP)
• Host: RAW264.7

Applications/Dilutions

• Dilutions:
  • In vitro assay
  • Ligand Activation
• Readout System:
  • Secreted Alkaline Phosphatase Reporter Assay Kit (Colorimetric) NBP2-25285

Packaging, Storage & Formulations

• Storage: Store in gas phase of liquid nitrogen.
• Reconstitution Instructions: Complete Growth Medium: DMEM with 4.5 g/L glucose + 10% FBS + 4 mM L-glutamine + 1 mM sodium pyruvate + 100 units/ml penicillin + 0.1 mg/ml streptomycin + 0.5 mg/ml G418 (Geneticin).

Details for Array

• Type: Cell

Notes

Assume all cultures are hazardous since they may harbor latent viruses or other organisms that are uncharacterized. The following safety precautions should be observed.
- Use pipette aids to prevent ingestion and keep aerosols down to a minimum.
- No eating, drinking or smoking while handling the RAW line.
- Wash hands after handling the RAW line and before leaving the lab.
- Decontaminate work surface with disinfectant or 70% ethanol before and after working with stable cells.
- All waste should be considered hazardous.
- Dispose of all liquid waste after each experiment and treat with bleach.

Alternate Names for Array

• SEAP assay

Limitations

This product is for research use only and is not approved for use in humans or in clinical diagnosis. Reporter Cell Lines are guaranteed for 1 year from date of receipt.

Publications for NFkB Secreted Alkaline Phosphatase Reporter Reporter Cell Line (NBP2-26261)(5)

Publications using NBP2-26261

• Zarpelon AC, Fattori V, Souto FO et al. The Sesquiterpene Lactone, Budlein A, Inhibits Antigen-Induced Arthritis in Mice: Role of NF-kB and Cytokines Inflammation 2017-08-05 [PMID: 28780730] (In vitro)
• Park Jh, Jun Jg, Kim Jk. (E)-3-(3,4-dihydroxy-2-methoxyphenyl)-1-(2,4-dihydroxyphenyl)prop-2-en-1-one, a novel licochalcone B derivative compound, suppresses lipopolysaccharide-stimulated inflammatory reactions in RAW264.7 cells and endotoxin shock in mice Chemico-Biological Interactions et al. 2014-12-05 [PMID: 25451593] (In vitro)
  
  Details:
  Mouse nFkB Secreted Alkaline Phosphatase Reporter SEAP - (SEAPorter(TM)) Stable Reporter Cell Line (also called RAW nF-kB/SEAP Reporter Cell Line ) used for nF-kB reporter assay - cells were pretreated or not with (E)-3-(3,4-dihydroxy-2-methoxyphenyl)-1-(2,4-dihydroxyphenyl)prop-2-en-1-one, a novel licochalcone B derivative compound and then were stimulated with 500 ng/ml LPS for 24 h (Fig. 4A)
• Namachivayam K, Blanco CL, Frost BL et al. Preterm human milk contains a large pool of latent TGF-B, which can be activated by exogenous neuraminidase. Am J Physiol Gastrointest Liver Physiol. 2013-06-15 [PMID: 23558011]
  
  Details:
  Products cited (Fig 5C): 1. RAW NF-kB/SEAP Reporter Cell Line (IML-120) 2. SEAPorter Assay Kit (10055K) Note: The results showed LPS activation of SEAP activity in the IML-120 cells could be attenuated by pre-treating the cells with pre-term milk + neuram
• Potzsch S, Blankenhorn A, Navarrete Santos A et al. The effect of an AGE-rich dietary extract on the activation of NF-kB depends on the cell model used. Food Funct. 2013-07-01 [PMID: 23426622]
  
  Details:
  IMGENEX products cited: 1. RAW NF-kB/SEAP Reporter Cell Line (IML-120): Figs 3, 4 2. SEAPorter Assay Kit (10055K): Fig 3 Note: Bread crumb extract (BCE) was used as an advanced glycation end product to treat RAW IML-120 cells. NF-kB activation was measure
• Feng D, Sangster-Guity N, Stone R et al. Differential requirement of histone acetylase and deacetylase activities for IRF5-mediated proinflammatory cytokine expression. J Immunol. 2010-11-15 [PMID: 20935208]